Bulbous Plant Species Research Articles

OPTIMIZATION OF THE MODE OF DEPOSITION OF ASEPTIC CULTURES IN VITRO OF A RARE AND ENDEMIC SPECIES ALLIUM IVASCZENKOAE

Optimization of the method of medium-term storage of the rare and endangered plant species Allium ivasczenkoae under in vitro conditions using modern approaches of biotechnology is an urgent task. Aseptic microplants Allium ivasczenkoae were used as starting material. The cultivation of microplants was carried out on nutrient media supplemented in various combinations with an increased concentration of sucrose, mannitol, chlorocholine chloride, ABA and BAP at various temperature and light conditions of deposition. The survival dynamics of A. ivasczenkoae varied depending on cultivation regimes. After 9 months of deposition, the survival of test-tube plants in mode 1 (control) was 20%, mode 2 - 60-80%, mode 3 - 30-50%. Under experimental conditions, the best results were obtained by depositing aseptic cultures for 9 months at a low positive temperature and low light intensity (mode 2) on nutrient media with the addition of CCC 120 mg/l and a sucrose concentration of 60 g/l and (B-1) and MS medium containing 0.5 mg/l BAP and 3 g/l mannitol (MSВМ). This mode of deposition made it possible to lengthen the period between transplants, as well as to maintain the viability of A. ivasczenkoae microplants from 60 to 80%. The data obtained make it possible to optimize the mode of medium-term storage of Allium ivasczenkoae under in vitro conditions for the mass production of rare species of bulbous plants in order to restore, reproduce and preserve the number of valuable gene pool and natural populations, create gene banks in vitro, reintroduce and green building.

Predicting the effect of climate change on the geographic distribution of the endemic Fritillaria aurea in Türkiye

Fritillaria aurea is a rare, high altitude, endemic, and bulbous plant species in Türkiye. Although it is classified as least concern according to IUCN criteria, the species has a narrow distribution. This study utilized ensemble modeling to forecast potential future changes in suitable habitats for F. aurea by two Shared Socio-Economic Pathways (SSPs: SSP 1-2.6 and 5-8.5). These pathways were constructed using two General Circulation Models (GCMs) and covered the years 2035, 2055, and 2085. The results showed that the minimum temperature of the coldest month (bio6), mean temperature of the wettest quarter (bio8), and precipitation of the warmest quarter (bio18) have the largest influence on the potential species distribution. The ensemble model predicted that the highly suitable habitats of F. aurea would contract under all future SSP scenarios and it would lose almost all of its potential highly suitable distribution areas by the end of the century. The remained population of F. aurea could possibly harbour in only minor areas of the North Anatolian Mountains in the north and Taurus Mountains in the south. The results of the study could contribute to establishing conservation strategies and natural resource management policies for F. aurea against the potential impacts of climate change. The highly suitable habitats under pessimistic scenarios at the end of this century projected by the present study can be determined as protected areas for the species.

Biological Activities of Galanthus fosteri Extracts: First Demonstration of the Interaction between Chlorogenic Acid and DNA Ligase by Molecular Docking.

Within the Amaryllidaceae family, the bulbous plant species Galanthus fosteri (G. fosteri) belongs to the Galanthus genus. Alkaloids with a broad variety of biological functions are typically found in the flora of this family. The G. fosteri plant's organs' antioxidant activity, antibacterial impact, and antimicrobial qualities were examined in this study. Total flavonoid contents (TFC) and total phenolic contents (TPC) of plant extracts were measured with spectrophotometric methods, and antioxidant activity was determined using the DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging technique. The HPLC method was used to determine the phenolic compounds on a component basis. The antibacterial properties of the extracts were assessed using the Kirby-Bauer disc diffusion method, and the minimum inhibitory concentration method against the pathogens Klebsiella pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli. Additionally, combination tests were performed between the extract and antibiotics. Leaf and stem extracts demonstrated greater antioxidant qualities than bulb extracts, despite the fact that extracts of plant organs did not exhibit appreciable levels of TPC, TFC, or antioxidant qualities. According to the HPLC analysis results, it was determined that chlorogenic acid was present in all of the extracts. In fact, it was determined that only chlorogenic acid was 8.02 (mg/10 g) in G. fosteri bulb peel, which has antimicrobial and antioxidant properties. A molecular docking study has demonstrated for the first time that the antibacterial effect of chlorogenic acid might be due to DNA replication inhibition.

A comprehensive evaluation of the chemical profiles and biological properties of six geophytes from Turkey: Sources of bioactive compounds for novel nutraceuticals

Geophytes are gaining interest as sources of natural ingredients in nutraceutical and pharmaceutical area. In this sense, six bulbous plant species from Turkey are investigated, namely Hyacinthella campanulata K.Perss. & Wendelbo, Muscari neglectum Guss. ex Ten., Tulipa humilis herb., Iris stenophylla Hausskn. ex Baker, Galanthus elwesii Hook. f. and Crocus danfordiae Maw. with the aim to highlight their chemical compositions and biological properties. Polyphenolic profiles of the different plant parts (flower, bulb and leaf) of the six genotypes were evaluated using colorimetric methods as well LC–MS/MS. The antioxidant properties and enzymes inhibitory potential (α-amylase, α-glucosidase, tyrosinase and cholinesterases) of the extracts were determined. Overall, highest total phenolic content and total flavonoid content were observed in the leaf extracts of the studied species, except for M. neglectum (flower extract) and T. humilis (flower extract). LC–MS/MS analysis revealed the abundance of some phenolic compounds including quinic acid, hesperidin and chlorogenic acid in selective extracts. The extracts showed significant antioxidant potentials, with leaf extract of the I. stenophylla being more potent, which is linked to its high phenolic contents. All the extracts displayed notable anti-acetylcholinesterase (1.77 – 2.53 mg GALAE/g) and tyrosinase (54.9–67.20 mg KAE/g). Selective extracts have showed activity against butyrylcholinesterase, with bulb extract of M. neglectum (2.99 mg GALAE/g), I. stenophylla (2.53 mg GALAE/g) and G. elwesii (2.52 mg GALAE/g) showing highest activity. Modest activity was observed against α-amylase and α-glucosidase. The experimental data gathered herein is the first report on the phytochemical and biological attributes of these bulbous plant species which project them as potential sources of biologically active compounds for phytomedicines and nutraceuticals development.

<p><strong><em>Trimelopter crispolanatum </em></strong><strong>(Asparagaceae, Scilloideae), a new species from Namaqualand, South Africa</strong></p>

Hyacinthaceae subfam. Ornithogaloideae (= tribe Ornithogaleae of Aparagaceae sensu APG 2009, 2016, Chase et al. 2009) includes about 300 species of bulbous plants mainly distributed throughout Europe, Africa and south-west Asia. The latest comprehensive study in Ornithogaloideae (Martínez-Azorín et al. 2011) demonstrates the existence of 19 monophyletic genera which are characterised by a clear syndrome of morphological characters related to biogeographic patterns.

Functional Diversity of Nectary Structure and Nectar Composition in the Genus Fritillaria (Liliaceae).

Fritillaria is a genus consisting of 130 to 140 species of bulbous plants, native to temperate regions of the northern hemisphere. Generally viewed as an insect pollinated genus with the exception of two North American species, Fritillaria gentneri and F. recurva, which are described as hummingbird-pollinated and the Asian species, F. imperialis, described as passerine-pollinated. These pollinator shifts are possibly the result of adaptive changes to the structure and morphology of the nectary, as well as a change in the nectar concentration and composition. A study was conducted in a target group of 56 Fritillaria species, based on the morphology of their nectaries and nectar composition to assess the significance of pollination mode as well as its predisposition for the evolution of bird pollination. All species studied had nectaries located at their tepal base and produced nectar, but their size, shape, color, and composition all varied. Most fritillaries had hexose-rich nectar, in easily accessible and unprotected nectaries. Scanning electron microscope (SEM) analysis revealed that the surface of the nectaries of most Fritillaria species was flat and clearly distinct from that of the surrounding tissues, which might be regarded as an adaptation for insect-pollination. Nectaries of F. imperialis were considerably larger and had dilute nectar without sucrose, which was produced profusely, thereby fulfilling the criteria characteristic of ornithophilous flowers. The copious nectar of presumed hummingbird-pollinated species was rather balanced and of medium sugar concentration. Their large lanceolate nectaries contrasted sharply with the tessellated background of their tepals. These characters might indicate a mixed pollination system that engages both birds and insects. Floral anatomy and microstructure and nectar composition for Fritillaria species in subgenera Korolkowia and Liliorhiza are studied for the first time.

Determining the factors affecting the distribution of Muscari latifolium, an endemic plant of Turkey, and a mapping species distribution model.

Species distribution modeling was used to determine factors among the large predictor candidate data set that affect the distribution of Muscari latifolium, an endemic bulbous plant species of Turkey, to quantify the relative importance of each factor and make a potential spatial distribution map of M. latifolium. Models were built using the Boosted Regression Trees method based on 35 presence and 70 absence records obtained through field sampling in the Gönen Dam watershed area of the Kazdağı Mountains in West Anatolia. Large candidate variables of monthly and seasonal climate, fine‐scale land surface, and geologic and biotic variables were simplified using a BRT simplifying procedure. Analyses performed on these resources, direct and indirect variables showed that there were 14 main factors that influence the species’ distribution. Five of the 14 most important variables influencing the distribution of the species are bedrock type, Quercus cerris density, precipitation during the wettest month, Pinus nigra density, and northness. These variables account for approximately 60% of the relative importance for determining the distribution of the species. Prediction performance was assessed by 10 random subsample data sets and gave a maximum the area under a receiver operating characteristic curve (AUC) value of 0.93 and an average AUC value of 0.8. This study provides a significant contribution to the knowledge of the habitat requirements and ecological characteristics of this species. The distribution of this species is explained by a combination of biotic and abiotic factors. Hence, using biotic interaction and fine‐scale land surface variables in species distribution models improved the accuracy and precision of the model. The knowledge of the relationships between distribution patterns and environmental factors and biotic interaction of M. latifolium can help develop a management and conservation strategy for this species.

Effect of plant age and vernalization on bolting, plant growth and enzyme activity of garlic (Allium sativum L.)

In many bulbous plant species that require vernalization, including garlic, the age of the plant and a period of low temperature are important for complete vernalization. However, the response of garlic to vernalization treatment, which regulates development, remains poorly understood. In this study, the effects of vernalization and plant age on garlic growth and development were assessed. Garlic cultivar cv. G064 (bolting type), which is widely cultivated in China, was grown in a plastic tunnel for three different durations (A65, A45 or A25: 65, 45 or 25 days after planting) and then exposed to four vernalization temperatures [T5–0: 5°C/0°C, T10–5: 10°C/5°C, T15–10: 15°C/10°C, T20–15: 20°C/15°C (day/night)] for 40 days in growth chambers. To evaluate the bolting rate, plant growth, and endogenous physiological indexes, leaf and pseudostem samples were taken from all plants after 20 days of treatment and from A25 plants after 40 days of treatment. The results indicate that the response to vernalization varied among garlic plants of different ages, although the patterns were similar among the treatments. In general, vernalization treatment significantly increased the bolting rate in plants at all ages tested, with the youngest plants having the highest bolting rates at 10°C/5°C (28.6%, 40.1% and 50.0% for the A65, A45, and A25 plants, respectively), inhibited vegetative growth (with reductions in plant height observed in all plants except for A65 plants, pseudostem diameter, and green leaf number, especially for the T5–10 treatment) and promoted peroxidase (E.C. 1.15.1.7) and superoxide dismutase (E.C. 1.15.1.1) activities compared with the results observed in the control. The lowest temperature induced the strongest enhancing effect. It seems that a curved parabolic relationship might exist between the vernalization conditions and successful vernalization. Furthermore, the regulation of growth and bolting of plants requiring green-plant vernalization is achievable through the control of plant age (sowing dates) and growth temperature.

Conservation biology of Chionodoxa lochiae and Scilla morrisii (Asparagaceae): Two priority bulbous plant species of the European Union in Cyprus

This paper presents data regarding conservation biology of Chionodoxa lochiae and Scilla morrisii; two threatened endemic plants of Cyprus, which are included as priority species in Annex II of the Habitats Directive. The population size and geographical distribution of the species were monitored for three years. C. lochiae was recorded in ten locations and S. morrisii was recorded in five locations. C. lochiae occurs in Pinus forests with/without Quercus alnifolia understory or in forest margins and riparian vegetation with Platanus orientalis. Favorable habitat of S. morrisii is the understory of Quercus infectoria stands and the Pistacia terebinthus-Quercus coccifera-Styrax officinalis shrubs. The distribution pattern of the species seems to follow habitat availability. Fecundity and Relative Reproductive Success of C. lochiae were stable and low, while in S. morrisii were constantly high. The lack of pollinators seems to be the main cause of the low sexual reproduction of C. lochiae. The germination strategy for both species is dependent on temperature. Some of the seeds are dormant and dormancy is broken by nitrates. The investigation of certain aspects of the biology of the two species yielded the information needed to identify the critical aspects affecting their survival and to propose sound conservation measures.

Hormonal regulation of gummosis and composition of gums from bulbs of hyacinth (Hyacinthus orientalis)

Hyacinth (Hyacinthus orientalis) bulbs infected by Fusarium oxysporum showed the symptoms of gummosis. The purpose of this study was to clarify the hormonal regulation of gummosis and composition of gums from hyacinth bulbs. The application of ethephon (2-chloroethylphosphonic acid), an ethylene-releasing compound, at 2% (w/w, in lanolin) induced gummosis in hyacinth bulbs. Methyl jasmonate (JA-Me) at 1.5% (w/w, in lanolin) induced gummosis as well. Simultaneous application of JA-Me and ethephon further enhanced gummosis. Molecular mass distribution of hyacinth gums analyzed by gel permeation chromatography indicated that the gums were mainly homogenous polysaccharides with an average molecular weight of ca. 30kDa. Analysis of the sugar composition of the gums after hydrolysis revealed that the majority were arabinose (ca. 35%) and galactose (ca. 40%) together with small amounts of fucose, rhamnose and uronic acids (ca. 5%, respectively), suggesting that the gums are pectic arabinogalactans. These results indicate that jasmonates (JAs) interact with ethylene to stimulate sugar metabolism, producing pectic arabinogalactans, and vice versa, leading to gummosis. These findings, together with those from our previous studies in tulips (Tulipa gesneriana) and grape hyacinth (Muscari armeniacum), revealed that sugar metabolism and hormonal regulation relating to gummosis are different among species of bulbous plants.

The effect of the ambient conditions on the life cycle of a bulbous plant

To evaluate how weather conditions affect the life cycle of a bulbous plant species, a methodology is proposed here using Crocus cambessedesii, a plant that is endemic to Mallorca and Menorca. Data from an experimental field study conducted from 2007 to 2013 in the south west of Mallorca is taken together with meteorological observations from Palma Airport, located about 10 km away from the site. The bias in the observations between the Airport and the field site is discussed, as well as the importance of performing meteorological measurements in situ to have a better knowledge of the local processes. It is found that flowering and fruiting periods start when the mean temperature is between the range of 16-20 C and 10-14 C, respectively. In addition, a certain amount of soil water content is needed (it has been raining during the previous days). These results are in agreement with those obtained for Crocus sativus, extensively studied for economic interests. Finally, some recommendations are made to apply this methodology to plants in other locations or other species.

Towards a Molecular Understanding of the Biosynthesis of Amaryllidaceae Alkaloids in Support of Their Expanding Medical Use

The alkaloids characteristically produced by the subfamily Amaryllidoideae of the Amaryllidaceae, bulbous plant species that include well know genera such as Narcissus (daffodils) and Galanthus (snowdrops), are a source of new pharmaceutical compounds. Presently, only the Amaryllidaceae alkaloid galanthamine, an acetylcholinesterase inhibitor used to treat symptoms of Alzheimer’s disease, is produced commercially as a drug from cultivated plants. However, several Amaryllidaceae alkaloids have shown great promise as anti-cancer drugs, but their further clinical development is restricted by their limited commercial availability. Amaryllidaceae species have a long history of cultivation and breeding as ornamental bulbs, and phytochemical research has focussed on the diversity in alkaloid content and composition. In contrast to the available pharmacological and phytochemical data, ecological, physiological and molecular aspects of the Amaryllidaceae and their alkaloids are much less explored and the identity of the alkaloid biosynthetic genes is presently unknown. An improved molecular understanding of Amaryllidaceae alkaloid biosynthesis would greatly benefit the rational design of breeding programs to produce cultivars optimised for the production of pharmaceutical compounds and enable biotechnology based approaches.

Transfer of knowledge about flowering and vegetative propagation from model species to bulbous plants

The extensive characterization of plant genes and genome sequences summed to the continuous development of biotechnology tools, has played a major role in understanding biological processes in plant model species. The challenge for the near future is to generate methods and pipelines for an efficient transfer of this knowledge to economically important crops and other plant species. In the case of flower bulbs, which are economically very important for the ornamental industry, flowering time control and vegetative propagation constitute the most relevant processes for agronomical improvements. Those processes have been reasonably studied in reference species, making them excellent candidates for translational investigations in bulbous plant species. The approaches that can be taken for the transfer of biological knowledge from model to non-model species can be roughly categorized as "bottom-up" or "top-down". The former approach usually goes from individual genes to systems, also known as a "gene-by-gene" approach. It assumes conservation of molecular pathways and therefore makes use of sequence homology searches to identify candidate genes. "Top-down" methodologies go from systems to genes, and are e.g. based on large scale transcriptome profiling via heterologous microarrays or RNA sequencing, followed by the identification of associations between phenotypes, genes, and gene expression patterns and levels. In this review, examples of the various knowledge-transfer approaches are provided and pros and cons are discussed. Due to the latest developments in transgenic research and next generation sequencing and the emerging of systems biology as a matured research field, transfer of knowledge concerning flowering time and vegetative propagation capacity in bulbous species are now within sight.

Seasonal variation in antimicrobial and phytochemical properties of frequently used medicinal bulbous plants from South Africa

The growing popularity of traditional medicine and the unrestricted collection of medicinal plants from the wild have put many of the slow growing bulbous plant species at the risk of over-exploitation and extinction in South Africa. This study was aimed at comparing the phytochemical composition and biological (antibacterial and anticandidal) activities of bulb and leaf extracts of Tulbaghia violacea, Hypoxis hemerocallidea, Drimia robusta and Merwilla plumbea between spring, summer, autumn and winter seasons, with the view of promoting the use of leaves, as a conservation strategy. Antibacterial and anticandidal activities of petroleum ether (PE), dichloromethane (DCM), 80% ethanol and water extracts of bulbs and leaves were tested against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae bacteria and the fungus Candida albicans using the microdilution bioassay. Spectrophotometric methods were used to evaluate saponin and phenolic compositions for the four seasons. Leaf and bulb extracts exhibited comparable anticandidal activity (MIC < 1 mg/ml) in all the plant species in all seasons. Only ethanol and water extracts of H. hemerocallidea corms (autumn and winter) showed correspondingly good fungicidal activity amongst the bulbs tested. Antibacterial activity was fairly comparable between bulbs and leaves with at least one extract of each plant species showing some good MIC values in most of the seasons. The best antimicrobial activities were recorded in winter and autumn seasons, with MIC values as low as 0.2 mg/ml in the DCM bulb extracts of T. violacea (winter) against K. pneumoniae and S. aureus. The amounts of total phenolic compounds in all plant samples were generally higher in spring compared to the other seasons. Condensed tannin, gallotannin and flavonoid levels, depending on the sample, were either higher in spring or winter except for H. hemerocallidea (corm) which had higher gallotannin levels in autumn. Total saponin levels were higher in winter in all plant samples. Although variation was observed in the phytochemical concentrations between the bulbs and leaves of each plant species, their antimicrobial activities were fairly comparable. Leaves may be used as substitutes for bulbs in the treatment of bacterial and fungal ailments.

Gummosis in grape hyacinth (Muscari armeniacum) bulbs: hormonal regulation and chemical composition of gums

The purpose of this study was to investigate the hormonal regulation of gummosis in grape hyacinth (Muscari armeniacum) bulbs, focusing especially on the chemical composition of the gums. The application of ethephon (2-chloroethylphosphonic acid), an ethylene-releasing compound, at 1% and 2% (w/w) in lanolin as well as ethylene induced gummosis in the bulbs within several days. Methyl jasmonate (JA-Me, 0.1-2% in lanolin) alone had no effect on gummosis. However, simultaneous application of JA-Me and ethephon led to extreme stimulation of ethephon-induced gummosis. Ethephon-induced gummosis in the bulbs depended on the maturation stage of the bulbs, increasing from April to July, but decreasing from August to September. Regardless of the presence of JA-Me, the application of ethephon to the inflorescence axis of grape hyacinths did not induce gummosis. Gel permeation chromatography analysis revealed that gums were homogenous polysaccharides with an average molecular mass of ca. 8.3 kDa. Analysis of the sugar composition of the gums after hydrolysis revealed that the molar ratio of Rha:Ara:Gal:GalA:GlcA was 25:10:40:7:15. These results suggest that principal factors of gummosis as well as the chemical composition of gums differ between species of bulbous plants.

GROWTH AND FLOWERING OF ORNITHOGALUM DUBIUM

GROWTH AND FLOWERING OF ORNITHOGALUM DUBIUM

Low temperature sensing in tulip (Tulipa gesneriana L.) is mediated through an increased response to auxin

Tulip (Tulipa gesneriana L.) is a bulbous plant species that requires a period of low temperature for proper growth and flowering. The mechanism of sensing the low temperature period is unknown. The study presented in this paper shows that the essential developmental change in tulip bulbs during cold treatment is an increase in sensitivity to the phytohormone auxin. This is demonstrated using a model system consisting of isolated internodes grown on tissue culture medium containing different combinations of the phytohormones auxin and gibberellin. Using mathematical modelling, equations taken from the field of enzyme kinetics were fitted through the data. By doing so it became apparent that longer periods of low temperature resulted in an increased maximum response at a lower auxin concentration. Besides the cold treatment, gibberellin also enhances the response to auxin in the internodes in this in vitro system. A working model describing the relationship between the cold requirement, gibberellin action and auxin sensitivity is put forward. Possible analogies with other cold-requiring processes such as vernalization and stratification, and the interaction of auxin and gibberellin in the stalk elongation process in other plant species are discussed.

The effect of temperature on seed germination in three common bulbous plants of different habitats in the Central Negev Desert of Israel

Seeds of three bulbous plant species of the Negev Desert of Israel were germinated in 10, 15, 20 and 25°C, which are temperatures commonly occurring in their natural habitats in winter when seed germination takes place. Germination rates and percentages correlated with their habitat distribution: Bellevalia desertorum, the most thermophilous species, germinated better and faster than the other species in all temperatures. The least thermophilous of the three, Tulipa systola, germinated very poorly in 20 and 25°C, while the intermediate B. eigii did very poorly in 25°C. Germination rates of the three species correlated with the maximum soil temperatures in the top 2-cm layer of the soil, and with the rate of drying of this soil layer in their respective habitats, as was measured one week after rainfall. High winter temperatures on south-facing slopes, B. desetorrum's habitat, and the associated rapid desiccation of the soil crust, may exclude T. systola and B. eigii from this habitat. However, low temperature is not a dominant factor causing the absence of B. desertorum or B. eigii from the coolest habitat, inhabited only by T. systola.

球根植物の耐凍性

Freezing tolerance of ornamental bulbs and corms of about 50 species or cultivars which were native to different climates in the world was assessed. Almost all of the bulbs and corms native to the South Africa could not survive freezing below -3 or -5°C. This suggests that the mild oceanic winters do not provide the stimulus to evolve hardy bulbous plant species. Hardy bulbous species native to Mediterranean coast, Asia Minor and East Asia survived freezing between -5°C and -23°C. Allium oreophilum, Fritillaria camtschatcensis and Lilium dauricum were the hardiest of the bulbs tested, tolerating temperatures of to -20°C to -23°C. In the hardy bulbs, the base plate was the least hardy plant part. The flower buds were also far less tolerant than the leaf and stem in many ornamental bulbous plants. Further study on freezing tolerance of flower buds in bulbous plants is required.